1.Effect of urine mixing degree on 24-hour urinary total protein in patients with chronic kidney disease based on generalized estimating equation
Shanfang QIU ; Weiwei WU ; Yang LI ; Yi FANG ; Xiaoyan JIAO ; Wenqi SHAO ; Shuan ZHAO ; Jie TENG ; Jieru CAI ; Xiaoqiang DING
Chinese Journal of Clinical Medicine 2024;31(3):428-432
Objective To explore the effect of urine mixing degree on 24-hour urinary total protein(24 h UTP)in patients with chronic kidney disease(CKD).Methods From October 1,2023 to December 31,2023,30 hospitalized patients who needed to complete 24 h UTP testing in Zhongshan Hospital,Fudan University were selected.A 5 L unified container was used to collect urine for 24 hours.After collection and one hour's standing,the urine sample was divided into upper,middle,and lower equal parts according to volume,which was defined as direct-sampling group.Then,the urine samples were fully mixed with a magnetic stirrer and sampled again according to the above-mentioned three-equal sampling method,which was defined as mixed-sampling group.The generalized estimating equation was used to compare the urinary protein concentration before and after mixing and at different sampling location.Results The results of generalized estimating equation showed that after controlling the variable"sampling position",there was no significant difference in urinary protein concentration between the direct-sampling group and the mixed-sampling group.After controlling the variable"mixing method",there was still no significant difference in urinary protein concentration at different sampling positions.After adjusting the covariates such as age,gender,and estimated glomerular filtration rate(eGFR),the results were consistent.Conclusions With standard protocol,the entire 24-hour urine sample is a relatively even-distributed solution.After the total urine collection is completed,the temporary sample can be directly extracted from any level of the original urine within 1 hour,and the urine protein concentration of the sample multiplied by the urine volume can reflect the 24 h UTR.
2.Introduction of a prospective cohort study of chronic and non-communicable diseases in general population in southernern China
Min XIA ; Minghuang HONG ; Xiaoqiang QIU ; Yingzi LIN ; Weisen ZHANG ; Peisong GAO ; Zhibin LI ; Zhijian HU
Chinese Journal of Epidemiology 2023;44(1):48-53
With the rapid changes in people's lifestyles, natural and social environments in recent years, the prevalence of chronic and non-communicable diseases in China and its related risk factors have also had tremendous changes. The epidemiological characteristics of chronic and non-communicable diseases and their risk factors vary throughout the country, and the impact of unique climate, diet and lifestyle in southern China on the incidence and prevalence of chronic and non-communicable chronic diseases remains to be elucidated. Therefore, large-scale cohort study is urgently needed to provide evidence for the etiological research and management of chronic and non-communicable chronic diseases in different areas, and for the national management strategy for major chronic and non-communicable diseases. The prospective cohort study of chronic and non-communicable diseases in general population in southern China was established in December 2017. The study recruited permanent residents aged 35-74 years from both urban and rural areas in Guangdong, Hainan, Fujian Provinces and Guangxi Zhuang Autonomous Region. A big data platform of precision medicine which integrates health information with biological samples for long-term follow up has been established. A baseline database of 116 520 people aged (54.9±12.5) years, including 71 077 women (61.0%), has been established. Collecting questionnaire survey data, physical examination data, and biological samples. This paper briefly introduces the concept, design and progress of the prospective cohort study of chronic and non-communicable diseases in general population in southern China.
3.Clinical efficacy of transanal total mesorectal excision on transanal endoscopic microsurgery platform in the treatment of middle and low rectal cancer
Xueshan BAI ; Guole LIN ; Xiaoqiang XUE ; Jiaolin ZHOU ; Junyang LU ; Huizhong QIU
Chinese Journal of Digestive Surgery 2021;20(3):339-345
Objective:To evaluate the clinical efficacy of transanal total mesorectal excision (taTME) on transanal endoscopic microsurgery (TEM) platform in the treatment of middle and low rectal cancer.Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of 28 patients with middle and low rectal cancer who underwent taTME on TEM platform in the Peking Union Medical College Hospital of Chinese Academy of Medical Science from October 2014 to October 2017 were collected. There were 21 males and 7 females, aged 59 years (51 years, 68 years). Observation indicators: (1) surgical and postoperative situations; (2) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect post-operative defecation function and survival of patients up to October 2020. Patients underwent physical examination, examination of tumor markers including carcinoembryonic antigen and CA19-9, colonoscopy, rectal magnetic resonance imaging, thoracoabdominal and pelvic enhanced computed tomography (CT) and (or) PET-CT examination during the follow-up. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the independent sample t test. Measurement data with skewed distribution were represented as M( P25,P75) or M (range), and comparison between groups was analyzed using the non parameter Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Results:(1) Surgical and postoperative situations: 28 patients underwent successful surgery, without intra-operative conversion to laparotomy. Of 28 patients, 24 cases underwent colorectal anastomosis and 4 cases underwent colon-anal anastomosis. Twenty-six cases underwent primary protective enterostomy and 2 cases didn't undergo primary protective enterostomy. The operation time of 28 patients was (182±37)minutes and the volume of intraoperative blood loss was 40mL(30 mL, 55 mL). One patient with intraoperative presacral hemorrhage received compression hemostasis. Eleven patients had postoperative complications, including 4 cases with anastomotic leakage, 2 cases with alteration of intestinal flora, 2 cases with paralytic ileus, 2 cases with urinary retention, 2 cases with urinary infection, 1 case with prolapse necrosis of small intestinal stoma, 1 case with anal hemorrhage, 1 case with rectovaginal fistula, 1 case with pelvic infection; some patients had multiple complications. Three patients had non-planned reoperation. One case without primary protective enterostomy had anastomotic leakage at postoperative 3 days, and was improved after emergency transversostomy. One case had prolapse necrosis of small intestinal stoma at postoperative 3 days and was improved after emergency enterostomy and reconstruction. One case with anal hemorrhage was stopped hemorrhage under anoscopy. Patients with other complications were cured after conservative treatments. The duration of postoperative hospital stay of 28 patients was 8 days(7 days, 9 days). Results of pathological examination in 28 patients showed 16 cases of moderately differentiated adenocarcinoma, 3 cases of moderately to highly differentiated adenocarcinoma, 5 cases of highly differentiated adenocarcinoma, 1 case of mucinous adenocarcinoma, 3 cases of pathological complete response. TNM staging of 28 patients showed 3 cases in stage T0N0, 4 cases in stage T1N0, 6 cases in stage T2N0, 4 cases in stage T2N1, 7 cases in stage T3N0, 3 cases in stage T3N1, 1 case in stage T4N1. The distance from tumor to distal margin was (2.2±1.7)cm. The surgical specimens of 28 patients showed negative for proximal, distal and circumferential margins. The number of lymph node dissection was 15±7. The complete rate of total mesorectal excision was 100%(28/28). Eleven of 28 patients underwent neoadjuvant therapy and 17 patients didn't receive neoadjuvant therapy. The tumor diameter, distance from tumor to anal margin, operation time, volume of intraoperative blood loss, duration of postoperative hospital stay were 2 cm(1 cm, 4 cm), 5 cm(4 cm, 6 cm), (187±25)minutes, 45 mL(38 mL, 53 mL), 8 days(7 days, 12 days) for patients with neoadjuvant therapy, respectively, versus 3 cm(2 cm, 4 cm), 5 cm(4 cm, 6 cm), (177±35)minutes, 40 mL(30 mL, 60 mL), 8 days(7 days, 8 days) for patients without neoadjuvant therapy, showing no significant difference between the two groups ( Z=-1.127, -0.293, t=0.590, Z=-0.790, -0.876, P>0.05). (2) Follow-up: 23 of 28 patients were followed up for (44±14)months. Of the 23 patients,11 cases were classified as grade A of Williams score for defecation function at postoperative 6 months, 8 cases were classified as grade B and 4 cases were classified as grade C. Eighteen of 23 patients with follow-up had disease-free survival, 1 of whom didn't undergo stoma closure due to anastomotic stenosis at postoperative 6 months. Three patients had distant metastasis, including 1 case with parastomal implantation metastasis, 1 case with sacral metastasis, 1 case with pulmonary metastasis. Two patients died, 1 case of whom died of urinary obstruction and 1 case with mucinous adenocarcinoma died at postoperative 24 months. Conclusion:TaTME based on TEM platform is feasible for middle and low rectal cancer, which has the advantages of preserving anus and negative circumferential margin.
4. An initial exploration of the application of transanal endoscopic microsurgery in rectal cancer patients with clinical complete response after neoadjuvant chemoradiotherapy
Xiaoqiang XUE ; Jiaolin ZHOU ; Guole LIN ; Xueshan BAI ; Yi XIAO ; Bin WU ; Huizhong QIU
Chinese Journal of Gastrointestinal Surgery 2019;22(6):560-565
Objective:
To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT).
Methods:
Using descriptive case series method, Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3-course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV-X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow-up results were used to evaluate the value of diagnosis and treatment of TEM on those patients.
Results:
There were 8 males and 5 females with a median age of 63 (27-80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0-7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0-7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42-70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5-30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (
5. Dose-response relationship between maternal hemoglobin concentration and preterm birth, in pregnant women
Liangjia WEI ; Shun LIU ; Dongping HUANG ; Xuefeng GUO ; Na WANG ; Yonghong SHENG ; Meile MO ; Jun LIANG ; Yanye SONG ; Xiaoqiang QIU
Chinese Journal of Epidemiology 2019;40(4):471-474
Objective:
To investigate the dose-response relationship between hemoglobin concentration and preterm birth, during pregnancy.
Methods:
With Zhuang ethnicity, a total of 12 780 pregnant women and their infants that admitted to Wuming、Pingguo、Jingxi、Debao、Longan and Tiandong hospitals, were recruited, in Guangxi Zhuang Autonomous Region, from January 2015 to December 2017. Non-conditional logistic regression method was used to analyze the effect of anemia on preterm birth during pregnancy. Dose-response relationship between hemoglobin concentration and preterm birth was explored, using the restrictive cubic spline model.
Results:
After excluding 2 053 pregnant women with hypertension or aged 35 years and over, results from the non-conditional logistic regression analysis showed that the risk of preterm birth in the anemia group was 1.29 times (
6. Associations of parental thalassemia with preterm birth and low birth weight
Xiaochun HUANG ; Xiaoqiang QIU ; Xiaoyun ZENG ; Shun LIU ; Fanglian WEI ; Yanning LI ; Tao LIU ; Yanan WU ; Baoying FENG ; Qunjiao JIANG ; Dongping HUANG
Chinese Journal of Epidemiology 2019;40(5):596-600
Objective:
To investigate the association between the preterm birth and low birth weight and parental thalassemia.
Methods:
Pregnant women and their husbands receiving prenatal examination in local hospitals or maternal and child health centers in Jingxi and Debao in Guangxi from January to December 2017 were selected as study subjects. A total of 758 pregnant women with pregnancy outcomes and their husbands, who were both or alone diagnosed with thalassemia through thalassemia gene detection, were selected as case group and 758 pregnant women with pregnancy outcomes and their husbands, who were negative in thalassemia gene detection and hemoglobin electrophoresis test were selected as control groups. The case group were further divided into mother group, father group and both mother and farther group. Clinical and pregnancy outcome data of the study subjects were collected for the analysis on the association between parental thalassaemia and preterm birth or low birth weight by the independent sample
7.Effect of lentivirus vector-mediated RNA interference dbpA gene silencing on the biological behavior of colorectal cancer cells
Ruiting LIU ; Yali HOU ; Xiangtian WU ; Guorong WANG ; Chang LIU ; Jirong BAI ; Jian QIU ; Likun YAN ; Xiaojun LI ; Xiaoqiang WANG
Chinese Journal of General Surgery 2019;34(7):613-617
Objective To investigate the effects of lentivirus-mediated RNA interference (RNAi) targeting DNA binding protein A (dbpA) on the proliferation and the biological behavior of colorectal cancer cell line SW620.Methods The experiment was divided into 3 groups:KD group (siRNA-dbpA,lentivirus interference group),CON group (non-specific sequence group) and NC group (blank control group).The lentiviral vector siRNA-dbpA was constructed and verified by PCR and DNA sequencing.SW620 cells were transfected with siRNA-dbpA plasmid,nontargeting siRNA plasmid,or empty plasmid.After 48 h the transfection,the cells were examined for dbpA expression using Western blot.After 72 hrs transfection,flow cytometry was used to detect the cell apoptosis and cell cycle changes.The cell growth inhibition rate was detected by MTT (4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay,and then clone formation was detected,and the ability of SW620 cells to form tumors in vivo after dbpA was silenced was studied in nude mice.Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting dbpA gene was successfully inserted into the lentiviral vector.siRNA-dbpA transfection resulted in reduced expression of dbpA in SW620 cells.After transfection,the apoptosis rate of siRNA-dbpA-transfected cells increased to 26.60% ± 0.38%,significantly higher than that in cells transfected with the nontargeting plasmid or the empty plasmid 12.54% ± 0.25% and 4.46% ± 0.19%,respectively (F =28.159,P <0.01).The growth inhibition test indicate that the OD value of the fifth day in siRNA-dbpA group was 0.194 ±0.037,significantly lower than that in the other two groups 0.814 ±0.043 and 1.625 ±0.061,respectively(F =23.214,P < 0.01).The colony formation number is 37 ± 3,64 ± 5and 175 ± 10 respectively,siRNA-dbpA is significantly higher than that in the other two groups(F =40.254,P < 0.01).After the completion of nude mouse transplantation tumor model,through the detection of tumor volume,KD group (group siRNA-dbpA) tumor volume after 14 d and CON and NC group had obvious difference (F =38.256,P < 0.05),and after 21d is more significant difference in tumor size (F =40.241,P < 0.01),can be clearly observed after 35 d KD group (group siRNA-dbpA) growing tumors had differences with the control group (F =30.257,P < 0.05).Conclusion Lentivirus-mediated RNAi targeting dbpA can effectively suppress the expression of dbpA in colorectal tumor in nude mice,it is proved that dbpA silencing has a significant inhibitory effect on the growth of living tumor cells and decrease the proliferation of the colorectal cells.
8.An initial exploration of the application of transanal endoscopic microsurgery in rectal cancer patients with clinical complete response after neoadjuvant chemoradiotherapy
Xiaoqiang XUE ; Jiaolin ZHOU ; Guole LIN ; Xueshan BAI ; Yi XIAO ; Bin WU ; Huizhong QIU
Chinese Journal of Gastrointestinal Surgery 2019;22(6):560-565
Objective To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT). Methods Using descriptive case series method,Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3?course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV?X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow?up results were used to evaluate the value of diagnosis and treatment of TEM on those patients. Results There were 8 males and 5 females with a median age of 63 (27?80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0?7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0?7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42?70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5?30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (n=5), which could be alleviated by non?steroidal anti?inflammatory drugs. One case had postoperative lung infection and was cured by antibiotic treatment. One case had urinary retention after removing urine catheter, and then a urine catheter was re?inserted. Average postoperative hospital stay was (2.8 ± 2.4) (2?12) days. All specimens were completely resected via TEM. Histopathological examination confirmed that 7 specimens had achieved pathologic complete response (pCR) and the other 6 specimens had obtained partial tumor response of CAP grade 2. Seven patients with pCR received a median follow?up of 24 (8?48) months and no local recurrence or distant metastasis was reported during follow?up period. Among these 7 cases, one developed defecation dysfunction after discharge, mainly for defecation pain and even dare to defecate, who returned to normal defecation within 2 months after surgery; One developed severe anal pain within six months after surgery and the pain disappeared after symptomatic pain relief. The other 6 patients with CAP grade 2 refused to undergo further radical operation because of their strong desire in preserving anus, and received remedial adjuvant chemotherapy instead. Conclusion For rectal cancer patients with cCR after nCRT, TEM does have certain application values if the patient has a strong desire to preserve anus.
9.An initial exploration of the application of transanal endoscopic microsurgery in rectal cancer patients with clinical complete response after neoadjuvant chemoradiotherapy
Xiaoqiang XUE ; Jiaolin ZHOU ; Guole LIN ; Xueshan BAI ; Yi XIAO ; Bin WU ; Huizhong QIU
Chinese Journal of Gastrointestinal Surgery 2019;22(6):560-565
Objective To preliminarily explore the value of transanal endoscopic microsurgery (TEM) in rectal cancer patients with clinical complete response (cCR) following neoadjuvant chemoradiotherapy (nCRT). Methods Using descriptive case series method,Clinical data of 13 patients who met the criteria of nCRT and were considered to be cCR after MRI or CT scanning, digital rectal examination and colonoscopic biopsy, as well as no lymph node or distant metastasis were found, then underwent TEM from 2013 to 2016 at the Department of General Surgery of Peking Union Medical College Hospital were collected retrospectively. A 3?course combination of capecitabine and oxaliplatin (XELOX) was used for chemotherapy. Besides, a 6MV?X ray radiation was used as radiotherapy simultaneously. Six to eight weeks after completion of radiotherapy, a preoperative assessment was carried out with intrarectal ultrasound, MRI, or pelvic abdominal CT examination. TEM was performed afterwards with informed consent. Postoperative pathological findings and follow?up results were used to evaluate the value of diagnosis and treatment of TEM on those patients. Results There were 8 males and 5 females with a median age of 63 (27?80) years. Preoperative examination showed that the lesions were located in the anterior wall in 3 cases, the posterior wall in 3 cases, the left side wall in 4 cases, and the right side wall in 3 cases. Before nCRT, the distance between tumor and anal margin was (4.8±1.1) (2.0?7.5) cm; after nCRT, this distance was (5.2±1.3) (3.0?7.5) cm. All the 13 patients underwent extended local resection of rectal cancer via TEM with the placement of urethral catheter. The average operative time was (52.2±3.7) (42?70) minutes, and the average intraoperative blood loss was (19.2±2.8) (5?30) ml. All the patients could engage in daily activities on postoperative day 1, and could cater themselves orally on postoperative day 2. The main discomfort was postoperative anal pain and foreign body sensation (n=5), which could be alleviated by non?steroidal anti?inflammatory drugs. One case had postoperative lung infection and was cured by antibiotic treatment. One case had urinary retention after removing urine catheter, and then a urine catheter was re?inserted. Average postoperative hospital stay was (2.8 ± 2.4) (2?12) days. All specimens were completely resected via TEM. Histopathological examination confirmed that 7 specimens had achieved pathologic complete response (pCR) and the other 6 specimens had obtained partial tumor response of CAP grade 2. Seven patients with pCR received a median follow?up of 24 (8?48) months and no local recurrence or distant metastasis was reported during follow?up period. Among these 7 cases, one developed defecation dysfunction after discharge, mainly for defecation pain and even dare to defecate, who returned to normal defecation within 2 months after surgery; One developed severe anal pain within six months after surgery and the pain disappeared after symptomatic pain relief. The other 6 patients with CAP grade 2 refused to undergo further radical operation because of their strong desire in preserving anus, and received remedial adjuvant chemotherapy instead. Conclusion For rectal cancer patients with cCR after nCRT, TEM does have certain application values if the patient has a strong desire to preserve anus.
10.Impact and changes of maternal hemoglobin on birth weight in pregnant women of Zhuang Nationality, in Guangxi
Jiehua CHEN ; Xuefeng GUO ; Shun LIU ; Jinghua LONG ; Guoqiang ZHANG ; Meichen HUANG ; Xiaoqiang QIU
Chinese Journal of Epidemiology 2017;38(2):154-157
Objective To investigate the hemoglobin (Hb) levels during pregnancy and Hb changes from early pregnancy and association with birth weight on infants.Methods Mothers of Zhuang Nationality who participated in the pregnancy care program and delivered at the Pingguo County Hospital from May 2013 to May 2015 were included in this study.Retrospective analysis was applied to collect data of health care and pregnancy outcomes.Multiple regression analysis and unconditional logistic regression model were used for data analysis.Results The mean birth weight appeared as (313 5.92± 435.84) grams.The Hb levels at early pregnancy showed significantly positive association with birth weight.Results from our study demonstrated that when Hb levels increased +1 g/dl at early pregnancy,birth weight would increase 17.61(95%CI:0.60-34.67) grams,in the adjusted model.The Hb levels at late pregnancy were significantly inversely associated with birth weight.Our findings suggested that when Hb levels increased + 1 g/di at late pregnancy,birth weight would reduce 19.61(95% CI:-37.53--1.70) grams in the adjusted model.Changes in Hb from early pregnancy stages were significantly inversely associated with birth weight after adjusting for confounders and Hb levels in the early pregnancy stages.The results also indicated that when Hb levels increased a + 1 g/dl from early to late pregnancy,the birth weight would decrease 32.63 g(95% CI:-48.93--16.32).Compared to the non-anemia group,the anemia group showed significantly increase of small-for-gestational-age (SGA)(OR=1.58,95%CI:1.08-2.32) in early pregnancy.Compared to women under the most reduction status,women with the least reduction had a significantly increase of SGA (OR=1.87,95%CI:1.24-2.81) among their infants.With the magnitude of reduction on Hb concentration during pregnancy,the risk of delivering babies with SGA showed a gradual trends of increase.Conclusion Hb levels at early pregnancy were positively associated with birth weight,but the changes of Hb were inversely associated with birth weight at late pregnancy,in women of Zhuang Nationality.Anemia in early pregnancy and the low amplitude of decreased Hb concentration during pregnancy were both risk factors for newborns under less gestational ages.

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