1.Renovascular morphological changes in a rabbit model of hydronephrosis.
Wan-Qiang, LI ; Zi-Qiang, DONG ; Xiao-Bing, ZHOU ; Bing, LONG ; Lu-Sheng, ZHANG ; Jian, YANG ; Xiao-Guang, ZHOU ; Ren-Ping, ZHENG ; Jie, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):575-81
Obstructive nephropathy ultimately leads to end-stage renal failure. Renovascular lesions are involved in various nephropathies, and most renal diseases have an ischemic component that underlies the resulting renal fibrosis. The aim of this study was to investigate whether morphological changes occur in the renal vasculature in hydronephrosis and the possible mechanisms involved. A model of complete unilateral ureteral obstruction (CUUO) was used. Experimental animals were divided into five groups: a normal control group (N) and groups of animals at 1st week (O1), 2nd week (O2), 4th week (O4) and 8th week (O8) after CUUO. Blood pressure was measured, renal arterial trees and glomeruli were assessed quantitatively, and renovascular three-dimensional reconstruction was performed on all groups. Glomerular ultrastructural changes were examined by transmission electron microscopy. The results showed that the systolic blood pressure was significantly increased in the obstructed groups (O1, O2, O4 and O8). Three-dimensional reconstruction showed sparse arterial trees in the O8 group, and a tortuous and sometimes ruptured glomerular basement membrane was found in the O4 and O8 groups. Furthermore, epithelial media thickness and media/lumen ratio were increased, lumen diameters were decreased, and the cross-sectional area of the media was unaltered in the segmental renal artery, interlobar artery and afferent arterioles, respectively. In conclusion, renal arterial trees and glomeruli were dramatically altered following CUUO and the changes may be partially ascribed to vascular remodeling. Elucidation of the molecular mechanisms of renovascular morphological alterations will enable the development of potential therapeutic approaches for hydronephrosis.
3.Protective effects of heat shock response on circulatory collapse induced by hyperthermia.
Bin WANG ; Bing-de LUO ; Fei ZOU ; Wei-ren WAN ; Jin-qiang GUO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(2):115-118
OBJECTIVETo investigate the protective effects and mechanism of heat shock response (HSR) on circulatory collapse induced by hyperthermia.
METHODSTwo experiments were carried out: (1) Protective effects of HSR. Rats were divided into 2 groups: heat shock (HS) group, sham control (SC) group. After HS group was pretreated with heat shock and recovered for 20 h at room temperature, both groups were exposed to heat till death, and blood pressure, electrocardiogram were measured continuously during exposure. Mean arterial pressure (MAP), survival time etc were acquired through Chart software. (2) Mechanism of effects. Rats were divided into 3 groups: HS group, SC group and normal control (NC) group. The treatment in HS and SC groups was identical with that in the first experiment, but it would be terminated at 73 min after heat exposure. Systolic pressure (Ps), diastolic pressure (Pd) etc were recorded and content of NO and HSP70 in myocardium were measured.
RESULTS(1) The survival time in HS group [(102.3 +/- 11.4) min] was longer than that in SC group [(87.9 +/- 7.7) min] and shock revealed later (P < 0.01); (2) During early heat exposure MAP in HS group was not different from that in SC group, but after 60 min MAP in HS group were higher than that in SC group; (3) MAP, Ps, Pd, HR and HSP70 in HS group were significantly higher but content of NO was lower than those in SC group (P < 0.01, P < 0.05).
CONCLUSIONHSR may induce upregulation of HSP70 and inhibit excessive production of NO in myocardium, thus result in relief of circulatory collapse induced by hyperthermia.
Animals ; Heat-Shock Proteins ; analysis ; Heat-Shock Response ; physiology ; Hot Temperature ; Male ; Nitric Oxide ; analysis ; Rats ; Rats, Sprague-Dawley ; Shock ; metabolism ; physiopathology ; Time Factors
4.Measurement of soluble CD44v6 in peripheral blood as assistant diagnosis of invasive pituitary adenomas.
Yan-guo KONG ; Chang-bao SU ; Zu-yuan REN ; Ren-zhi WANG ; Gui-lin LI ; Wan-chen DOU ; Bo ZHANG ; Shi-qiang TIAN
Acta Academiae Medicinae Sinicae 2003;25(6):698-701
OBJECTIVETo investigate the value of measuring the concentration of soluble CD44 splice variant 6 (sCD44v6) in peripheral blood in patients with invasive and non-invasive pituitary adenomas.
METHODSThe concentrations of sCD44v6 in peripheral blood were measured with ELISA in 68 patients with invasive pituitary adenomas and 100 patients with non-invasive pituitary adenomas.
RESULTSThe serum concentration of sCD44v6 in patients with invasive pituitary adenomas was lower than that in patients with non-invasive pituitary adenomas, while the latter was lower than that in healthy controls. The serum concentrations of sCD44v6 were (44.63 +/- 7.21), (34.53 +/- 6.41), and (26.34 +/- 4.95) ng/ml in patients with invasive microadenoma, macroadenoma, and giant adenoma, and (60.78 +/- 9.61), (57.78 +/- 10.00), and (37.22 +/- 5.17) ng/ml in patients with non-invasive microadenoma, macroadenoma, and giant adenoma, lower than that in the healthy control group (68.73 +/- 6.00) ng/ml. Significant differences were observed among groups (P < 0.005). The concentration of sCD44v6 in peripheral blood decreased as the tumor size increased (P < 0.01), which was particularly significant in invasive pituitary adenomas. The positive rate in the patients with invasive pituitary adenomas reached 89.71%.
CONCLUSIONSerum concentration of sCD44v6 in the peripheral blood is inversely correlated with tumor size and its invasive growth, which may provide certain value in the early diagnosis, treatment and prognosis of invasive pituitary macroadenoma and giant adenoma.
Adenoma ; blood ; diagnosis ; pathology ; Adult ; Biomarkers, Tumor ; Female ; Glycoproteins ; blood ; Humans ; Hyaluronan Receptors ; blood ; Male ; Middle Aged ; Neoplasm Invasiveness ; Pituitary Neoplasms ; blood ; diagnosis ; pathology ; Prognosis
5.Renovascular morphological changes in a rabbit model of hydronephrosis.
Wan-qiang LI ; Zi-qiang DONG ; Xiao-bing ZHOU ; Bing LONG ; Lu-sheng ZHANG ; Jian YANG ; Xiao-guang ZHOU ; Ren-ping ZHENG ; Jie ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):575-581
Obstructive nephropathy ultimately leads to end-stage renal failure. Renovascular lesions are involved in various nephropathies, and most renal diseases have an ischemic component that underlies the resulting renal fibrosis. The aim of this study was to investigate whether morphological changes occur in the renal vasculature in hydronephrosis and the possible mechanisms involved. A model of complete unilateral ureteral obstruction (CUUO) was used. Experimental animals were divided into five groups: a normal control group (N) and groups of animals at 1st week (O1), 2nd week (O2), 4th week (O4) and 8th week (O8) after CUUO. Blood pressure was measured, renal arterial trees and glomeruli were assessed quantitatively, and renovascular three-dimensional reconstruction was performed on all groups. Glomerular ultrastructural changes were examined by transmission electron microscopy. The results showed that the systolic blood pressure was significantly increased in the obstructed groups (O1, O2, O4 and O8). Three-dimensional reconstruction showed sparse arterial trees in the O8 group, and a tortuous and sometimes ruptured glomerular basement membrane was found in the O4 and O8 groups. Furthermore, epithelial media thickness and media/lumen ratio were increased, lumen diameters were decreased, and the cross-sectional area of the media was unaltered in the segmental renal artery, interlobar artery and afferent arterioles, respectively. In conclusion, renal arterial trees and glomeruli were dramatically altered following CUUO and the changes may be partially ascribed to vascular remodeling. Elucidation of the molecular mechanisms of renovascular morphological alterations will enable the development of potential therapeutic approaches for hydronephrosis.
Animals
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Blood Pressure
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Disease Models, Animal
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Glomerular Basement Membrane
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blood supply
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pathology
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physiopathology
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Hydronephrosis
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pathology
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physiopathology
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Male
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Rabbits
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Renal Artery
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pathology
;
physiopathology
6.Cloning of CTB-PROIN fusion gene and its expression in Escherichia coli.
Li CHEN ; Feng-Xiu OUYANG ; Bing-Jun QIAN ; Hong REN ; Qiang WANG ; Qing-Wu JIANG ; Yu-Jiong WANG ; Jing-Bo LIU ; Wan-Qi LIANG ; Da-Bing ZHANG
Chinese Journal of Biotechnology 2005;21(2):204-210
A fusion gene CTB-PROIN, in which Proinsulin gene was fused to the 3' end of CTB gene by a hinge peptide 'GPGP', was constructed and cloned into pET-30a(+) to obtain a prokaryotic expression vector pETCPI. Subsequently the recombinant plasmid pETCPI was transformed into E. coli stain BL21 (DE3). After induced by IPTG, the expression product was analyzed by sodium dodecyl sulphate-polyacrylamide gel (15%) electrophoresis (SDS-PAGE), and its result indicated that the recombinant protein CTB-PROIN was expressed and accumulated as inclusion bodies. The recombinant CTB-PROIN protein accumulated to the level of 25% of total bacterial proteins. After inclusion bodies was denaturalized and refolded in vitro, significant assembly of monomers had occurred, and the recombinant protein represented assembled pentamers. The results of western blotting analysis also demonstrated that the fusion protein could be recognized by the anti-CT and anti-insulin antibody, respectively. In addition, the result of the CTB-PROIN-GM1 binding assay, that the protein could bind to monosialoganglioside specifically, showed it possesed biological activity in vitro. These results provided the possibility of developing a cheaper and more efficient oral vaccine for type I diabetes using such constructs.
Artificial Gene Fusion
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Cholera Toxin
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genetics
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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G(M1) Ganglioside
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metabolism
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Proinsulin
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genetics
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Recombinant Proteins
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biosynthesis
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genetics
7.Effect of Epstein-Barr virus nuclear antigen 1 on cell proliferation and cell cycle in nasopharyngeal carcinoma cells.
Ren-qiang WAN ; Xiang-ping LI ; Lu WANG ; Juan LU ; Hua-nan LUO ; Gang LI ; Xiong LIU ; Bao ZHANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(8):675-680
OBJECTIVETo study the effect of Epstein-Barr virus nuclear antigen 1 (EBNA1) on cell proliferation and cell cycle in nasopharyngeal carcinoma (NPC) cells.
METHODSRecombinant lentivirus that encoded EBNA1 short hairpin RNA (shRNA) was prepared. The C666-EBNA1 (CE) cells were transduced with lentivirus and selected by fluorescence activated cell sorting (FACS) to repress EBNA1 expression. The protein expression levels of EBNA1 were examined by Western blot. The effect of EBNA1 silence on cell proliferation was analyzed by MTT assay and cell growth assay, respectively. Cell cycle was assessed by flow cytometry. The mRNA and protein levels of cell cycle regulators were examined by real-time PCR and Western blot.
RESULTSRecombinant lentivirus encoded EBNA1 shRNA was successfully constructed. The EBNA1 expression in CE cells was significantly reduced by lentivirus-mediated RNA interference. The results of cell counting and MTT assay showed that EBNA1 down-regulation significantly inhibited cell growth in CE-shRNA EBNA1 cells (P < 0.05). Compared with the control group, the percentage of cells in G0-G1 phase was increased from (62.43 ± 6.62)% to (89.66 ± 0.64)% (t = -7.091, P = 0.002), and that in S phase was decreased from (34.93 ± 7.36)% to (7.82 ± 2.44)% (t = 6.095, P = 0.004). The mRNA expressions of c-myc, CDK4, CDK6 and pRb were decreased by 65.60%, 34.06%, 41.05% and 55.29% respectively with the similar results in protein expression levels.
CONCLUSIONSSuppression of EBNA1 may inhibit the growth of nasopharyngeal carcinoma cells in vitro and induce a G1-phase cell cycle arrest, which might be mediated by down-regulation of c-myc, CDK4, CDK6 and pRb.
Carcinoma ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Epstein-Barr Virus Nuclear Antigens ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Lentivirus ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transduction, Genetic
8.Inhibitory effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer.
Zhi-wei ZHOU ; De-sen WAN ; Guo-qiang WANG ; Jing-qing REN ; Zhen-hai LU ; Shao-xian TANG ; Yan-li YE ; Gong CHEN ; Su-xia LIN
Chinese Journal of Gastrointestinal Surgery 2006;9(2):161-164
OBJECTIVETo study the effect of angiogenesis inhibitor YH-16 in combination with 5-FU on liver metastasis of colorectal cancer.
METHODSIn vitro, the inhibitory effects of YH-16 and 5-FU on the growth of vascular endothelial cells and colorectal cancer cells were examined by MTT assay. In vivo, colorectal cancer cells were transplanted into BALB/c mice, and the mice were divided into six groups randomly:control group, low-dose YH-16 group, middle-dose YH-16 group, high-dose YH-16 group, 5-FU group and combination group. The number of liver metastases, the size of primary tumor and the toxicity were examined after 2 weeks postoperatively. The expression of vascular endothelial growth factor (VEGF) in liver metastases was detected by immunohistochemistry, and tumor microvessel density (MVD) was measured by immunostaining with CD34 and factor VIII (monoclonal antibodies.
RESULTSIn vitro, YH-16 inhibited the growth of colon cancer cells and vascular endothelial cells, with the IC50 at (2.16+/-0.28) microg/ml and (0.64+/-0.10) microg/ml respectively. In vivo high-dose YH-16 and 5-FU had a remarkable inhibitory effect on liver metastasis, and the combination group showed significant enhancement on this effect (P< 0.05). The combination group and 5-FU group could inhibit the growth of primary tumor, but not found in YH-16 group. The toxicity of YH-16 was lower than that of 5-FU (P< 0.05), and the difference was not found in the toxicity between combination group and 5-FU group (P > 0.05). Expression of VEGF in liver metastases was clearly inhibited by YH-16 in combination with 5-FU or 5-FU alone compared to the control group, and MVD in middle-dose and high-dose YH-16 group, 5-FU group and combination group was lower than that in control group (P< 0.05).
CONCLUSIONSThe angiogenesis inhibitor YH-16 can inhibit liver metastasis of colorectal cancer through inhibiting the growth of vascular endothelial cells. YH-16 in combination with 5-FU has additive effect on inhibitory activity against liver metastasis.
Angiogenesis Inhibitors ; therapeutic use ; Animals ; Cell Line, Tumor ; Colorectal Neoplasms ; drug therapy ; pathology ; Drug Therapy, Combination ; Female ; Fluorouracil ; therapeutic use ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor A ; metabolism
9.The mechanism of signal extension in Haoqin Qingdan decoction immunity activity in Damp-heat syndrome of pneumonia disease infected by influenza virus.
Yuan PAN ; Bing-de LUO ; Pei-zheng LIN ; Ye LIU ; Wei-ren WAN ; Jin-qiang GUO
Chinese Journal of Preventive Medicine 2010;44(7):612-616
OBJECTIVETo explore the immunoregulation existing signal transduction mechanism, to evaluate the role of lay its experimental basis By using Haoqin Qingdan decoction for treatments on the mouse models.
METHODSA total of 40 NIH Mice were randomly divided into five groups: control group, virus group (infecting by influenza virus), complex model group (richly fatty and sweet diet + Humid heat environment + infecting by influenza virus), virazole group (mouse of model group was treated by virazole), and Haoqin Qingdan decoction group (mouse of complex model group was treated by decoction of Haoqin Qingdan). When the complex model was established, determination of the mice lung indexes in each group and calculate the inhibition of lung indexes. The level of TLR2 mRNA and NF-κB mRNA expressions of peritoneal macrophages in each group of mice were quantitated by reverse transcription-polymerase chain reaction (RT-PCR). The level of IL-4 and IFN-γ in mouse serum was detected by ELISA to calculate the Th1/Th2 (IFN-γ/IL-4).
RESULTSThe lung index of control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were separately: (0.79 ± 0.11)%, (1.93 ± 0.38)%, (1.41 ± 0.26)%, (1.10 ± 0.26)% and (1.02 ± 0.16)%; The mice of virazole group and Haoqin Qingdan decoction group lung index were decreased (t = 0.322, P < 0.05). TLR2 mRNA expression The results showed that the control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were: 0.145 ± 0.017, 0.991 ± 0.149, 0.903 ± 0.124, 0.257 ± 0.03 and 0.413 ± 0.031; Compared to the complex model group, Haoqin Qingdan decoction group and virazole group were decreased (t = 0.422, F = 112.834, P < 0.05). Control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group NF-κB mRNA expression were separately: 0.075 ± 0.148, 0.379 ± 0.019, 0.291 ± 0.012, 0.169 ± 0.026 and 0.175 ± 0.033; the expression in virazole group and Haoqin Qingdan decoction group were decreased (t = 0.422, F = 112.834, P < 0.05). The level of IFN-γ in mice serum of control group, virus group, complex model group, virazole group and Haoqin Qingdan decoction group were: (7434.06 ± 323.27) pg/ml, (8679.77 ± 198.70) pg/ml, (8068.78 ± 113.8) pg/ml, (7454.66 ± 301.30) pg/ml and (7484.56 ± 229.85) pg/ml respectively; the IFN-γ level in serum of Haoqin Qingdan decoction group and virazole group were decreased (t = 0.201, F = 5.390, P < 0.05). Each group of mice IL-4 contents were (3701.74 ± 256.00) pg/ml, (3569.64 ± 161.35) pg/ml, (3530.88 ± 334.63) pg/ml, (3481.84 ± 282.25) pg/ml and (3618.00 ± 262.16) pg/ml; there were no significant difference between each group (t = 0.414, F = 0.505, P > 0.05). Th1/Th2 type cells in state of equilibrium (means IFN-γ/IL-4) were: 2.02 ± 0.19, 2.38 ± 0.10, 2.36 ± 0.14, 2.22 ± 0.17 and 2.07 ± 0.15; and complex model group Haoqin Qingdan decoction group and virazole group were decreased, and there was no significant difference observed (t = 0.587, F = 3.684, P > 0.05).
CONCLUSIONThe effect of Haoqin Qingdan decoction on treatment of damp-heat syndrome of pneumonia infected by influenza virus was observed. Through reducing the expressions of TLR2, it decreases the levels of NF-κB mRNA and the proportionality of Th1/Th2 are obviously descend (P < 0.05). Haoqin Qingdan decoction can reduce the lung index and relieve the pathogenic changes.
Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Lung ; pathology ; virology ; Male ; Mice ; Mice, Inbred Strains ; NF-kappa B ; immunology ; Orthomyxoviridae ; pathogenicity ; Phytotherapy ; Pneumonia, Viral ; drug therapy ; immunology ; virology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Toll-Like Receptor 2 ; immunology
10.Analysis on the mortality of colorectal and anal cancer in China during 2004 - 2005.
Ming WU ; Si-Wei ZHANG ; Ren-Qiang HAN ; Jin-Yi ZHOU ; Xiao-Nong ZOU ; Wan-Qing CHEN
Chinese Journal of Preventive Medicine 2010;44(5):403-407
OBJECTIVETo describe the mortality of colorectal and anal cancer in the Chinese population during 2004 - 2005.
METHODSMortality of colorectal and anal cancer from The 3rd National Death Retrospective Sampling Survey (2004 - 2005) were analyzed, with that the total population was 142 660 482 person-year and the number of death cases was 10 586. Crude death rate, age-standardized death rate by Chinese standard population (CASR) and world standard population (WASR), the constitute proportion to all cancer deaths and rank of cancer death were calculated and compared with The 1st (during 1973 - 1975) and The 2nd (during 1990 - 1992) National Death Retrospective Surveys.
RESULTSThe mortality of colorectal and anal cancer in China was 7.42/100 000 (10 586/142 660 482) during 2004 - 2005, accounting for 5.46% of total cancer deaths and ranked the 5th leading cause of death from cancer. CASR and WASR were 4.79/100 000 and 6.57/100 000, respectively. Gender specific mortality was higher for males with 8.38/100 000 (6114/72 970 241) than for females with 6.42/100 000 (4472/69 690 241). The crude death rates were 10.01/100 000 (4796/47 899 806) in urban areas and 6.11/100 000 (5790/94 760 676) in rural areas, moreover, the crude death rates in Eastern, Middle and Western part of China were 8.67/100 000 (4558/52 556 694), 7.19/100 000 (3580/49 781 225) and 6.07/100 000 (2448/40 322 563) respectively. Compared to the crude death rate 5.30/100 000 and CASR 4.54/100 000 during 1990 - 1992, the crude death rate and CASR from colorectal and anal cancer increased by 40.00% and 5.51%, whereas compared to the crude death rate 4.17/100 000 and CASR 4.27/100 000 during 1973 - 1975, the crude death rate and CASR had increased by 77.94% and 12.18% respectively.
CONCLUSIONThe mortality of colorectal and anal cancer has been increasing rapidly in China. The mortality is higher in males, and appears to be diverse in different areas.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anus Neoplasms ; epidemiology ; mortality ; Cause of Death ; Child ; Child, Preschool ; China ; epidemiology ; Colorectal Neoplasms ; epidemiology ; mortality ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Survival Rate ; Young Adult