Objective To explore the effect of berberine on the proliferation and invasion of human colon cancer cells and its mechanism.Methods The paraffin-embedded tumor tissues and intestinal incisal tissues(control tissues)of 96 patients who underwent radical resection of colon cancer at the First Affiliated Hospital of Henan University from January 2021 to December 2022 were selected as the research subjects.The expression of hypoxia-inducible factor-1α(HIF-1α)in tumor tissues and control tissues was detected by using the immunohistochemical method,and the relationship between HIF-1α and clinicopathological features was analyzed.The human colon cancer cell line HCT116 was cultivated in normoxic and hypoxic conditions,respectively;after adhesion,the cells were divided into normoxic group,hypoxic group,normoxic combined with berberine group,and hypoxic combined with berberine group.The cells in the normoxic combined with berberine group and the hypoxic combined with berberine group were treated with 25,50,75,and 100 μmol·L-1 berberine,respectively;while the cells in the normoxic group and the hypoxic group were not treated with any drugs.At 24,48,72,and 96 hours of cultivation,the proliferation of cells in each group was detected by using the thiazolyl blue tetrazolium bromide method,respectively.Based on the above results,the most suitable drug concentration was determined as 75 μmol·L-1.At 24 hours of cultivation,the invasion of cells in the normoxic group,normoxic combined with 75 μmol·L-1 berberine group,hypoxic group,and hypoxic combined with 75 μmol·L-1 berberine group was detected by using the Transwell method;at 48 hours of cultivation,the relative expression levels of HIF-1α and vascular endothelial growth factor(VEGF)proteins in the cells in each group were detected by using Western blot.Results HIF-1α was mainly expressed in the nucleus of colon cancer tissues and control tissues.The positive expression rates of HIF-1α in control tissues and colon cancer tissues were 19.8％(19/96)and 67.7％(65/96),respectively;the positive expression rate of HIF-1α in colon cancer tissues was significantly higher than that in control tissues(x2=11.298,P＜0.05).The expression of HIF-1α in colon cancer tissues was correlated with tumor size,lymph node metastasis,and TNM staging(P＜0.05),but had no correlation with patient gender,age,tumor infiltration depth,and differentiation degree(P＞0.05).The cells in the normoxic group showed logarithmic growth after 24 hours of culture;and the addition of berberine at a concentration of 25 μmol·L-1 had no significant effect on cell proliferation(P＞0.05);compared with 0 μmol·L-1berberine,the addition of berberine at concentrations of 50 μmol·L-1 and 75 μmol·L-1 significantly reduced cell proliferation activity after 48 hours of culture(P＜0.05),and the addition of berberine at a concentration of 100 μmol·L-1 significantly reduced cell proliferation activity after 24 hours of culture(P＜0.05).After 48 hours of culture with berberine at a concentration of 25 μmol·L-1,the proliferation activity of cells in the hypoxic group was significantly higher than that in the 0 μmol·L-1 berberine(P＜0.05);compared with 0 μmol·L-1 berberine,the addition of berberine at concentrations of 50,75,and 100 μmol·L-1 significantly reduced cell proliferation activity after 24 hours of culture(P＜0.05),and the higher the concentration of berberine,the more pronounced the decrease in cell proliferation activity(P＜0.05).At the same concentration of berberine,compared with the normoxic group,the hypoxic group showed a more significant decrease in cell proliferation activity and a more pronounced slowing-down trend in cell proliferation(P＜0.05).There was no statistically significant difference in the number of cells crossing the membrane between the normoxic group and the normoxic combined with 75 μmol·L-1 berberine group(t=0.955,P＞0.05);the number of cells crossing the membrane in the hypoxic combined with 75 μmol·L-1 berberine group was significantly less than that in the hypoxic group(t=5.354,P＜0.05);the number of cells crossing the membrane in the hypoxic group was significantly more than that in the normoxic group(t=2.625,P＜0.05).There was no statistically significant difference in the relative expression levels of HIF-1α and VEGF proteins between the normoxic group and the normoxic combined with 75 μmol·L-1 berberine group(P＞0.05);the relative expression levels of HIF-1α and VEGF proteins in the hypoxic group were significantly higher than those in the normoxic group(P＜0.05);the relative expression levels of HIF-1α and VEGF proteins in the hypoxic combined with 75 μmol·L-1 berberine group were significantly lower than those in the hypoxic group(P＜0.05);there was no statistically significant difference in the relative expression levels of HIF-1α and VEGF proteins between the normoxic combined with 75 μmol·L-1 berberine group and the hypoxic combined with 75 μmol·L-1 berberine group(P＞0.05).Conclusion HIF-1α expression is associated with growth and invasion of colon cancer.Berberine may inhibit the proliferation and invasion of colon cancer cells HCT116 by regulating the expression of HIF-1α and its downstream factor VEGF.

Objective:To explore the efficacy and safety of single-incision transobturator bulbourethral sling suspension without skin tunnel puncture in male patients with urinary incontinence.Methods:The clinical data of 6 male patients with urinary incontinence who underwent single-incision transobturator bulbourethral sling suspension without skin tunnel puncture in Beijing Hospital from August 2023 to August 2024 were retrospectively analyzed.The age of the patients ranged from 66 to 76 years old, with an average of 71.7 years old. The disease duration ranged from 18 to 48 months, with an average of 30 months. Six patients used 1 to 3 pads per day, with an average of 2.3 pads. The International Continence Incontinence Questionnaire Short Form (ICI-Q-SF) scored 13 to 19, with an average of 15.8. The Incontinence Quality of Life Questionnaire (I-QOL) scored 5.3 to 30.6, with an average of 18.8. Three patients underwent transurethral resection of the prostate for benign prostatic hyperplasia and three patients underwent radical prostatectomy for prostate cancer. The degree of urinary incontinence was mild in 2 cases and moderate in 4 cases. The technical points are as follows: the puncture method has been changed from the traditional outside-in approach to an inside-out approach. After the puncture needle passes through from beneath the skin at the incision, the sling is guided in, avoiding the need for skin tunneling punctures. Upon completion of the puncture, the ends of the sling on both sides are tied with a certain tension at the midline of the incision, and the incision is then closed layer by layer. The efficacy and safety of surgery were evaluated by recording the number of daily pad use, subjective scoring scale [International Committee on Urinary Incontinence Questionnaire-Short Form (ICI-Q-SF), Incontinence Quality of Life (I-QOL)] and complications at 1 month after surgery. Social continence was defined as 0 to 1 pad use per day. Successful treatment was defined as social continence. Treatment improvement was defined as no social continence, but 50% or more improvement of symptoms compared with that before surgery. Other conditions were defined as treatment failure.Results:All operations were successfully completed. After 1 to 11 months of follow-up, all patients achieved social continence. The patients' postoperative daily use of urinary pads ranged from 0 to 1 piece, with a mean of 0.5 piece. ICI-Q-SF scores ranged from 1 to 7, with a mean of 3. I-QOL scores ranged from 72.1 to 85.2, with a mean of 77.0. All the indicators were significantly improved compared with those before operation. In terms of postoperative complications, one patient had dysuria and urinary retention 2 days after the removal of the catheter, which was improved after symptomatic treatment of anti-inflammatory, detumescence, and indwelling catheter. At the last follow-up, there were no surgical related complications.Conclusions:The single-incision transobturator bulbourethral sling suspension without skin tunnel puncture for the treatment of male urinary incontinence is safe and effective. Compared to the traditional surgical method, it does not increase the difficulty of the procedure and is technically feasible, offering clinicians a new approach and perspective.

Objective:To evaluate the clinical value of nanopore targeted sequencing (NTS) in pathogens detection in urinary tract by comparing the results of different tests performed on the same urine sample.Methods:The results of NTS and urine culture test collected from 326 patients in the Department of Urology of People's Hospital of Wuhan University from July 2020 to June 2021 were retrospectively analyzed. There were 224 males and 102 females. The average age was (56.88 ± 14.58)years old. χ 2 test and Student’s test and Wilcoxon's sign rank test were used to analyze the differences of the pathogen detection rate, pathogen types results and detection time consuming between NTS and urine culture. The clinical value of the NTS in rapid detection of urinary pathogens was evaluated. Results:Among 326 hospitalized patients, the urinary tract microbes’ detecting rate of NTS was significantly higher than that of urine culture[67.80%(221/326)vs. 23.93%(78/326), χ2=130.25, P<0.01]. The uropathogens detecting rate of NTS was significantly higher than that of urine culture[54.29%(177/326)vs. 23.31%(76/326), χ2=38.95, P<0.01]. The number of urinary tract microbes detected by NTS was significantly higher than that of urine culture ( Z=11.49, P<0.01), the number of uropathogens was significantly higher than that of urine culture ( Z=9.67, P<0.01). The detection time of NTS and urine culture positive samples was (24.29±2.65) h and (49.28±11.30) h, the difference was statistically significant ( t =39.48, P<0.01). The results obtained by using NTS and urine culture were consistent in 135 (41.41%) samples. In 150 (46.01%) samples, NTS could detect the urinary tract microbes while urine culture cannot find, of which 112 cases (34.36%) were uropathogenic. In 27 cases (8.28%), more pathogens were detected by NTS except those from urine culture. In 6 cases (1.84%) re-detecting NTS after antibiotic therapy, the number of reads of primary uropathogen decreased gradually with the growth of colonizing bacteria or opportunistic pathogens appeared in the end. Re-examinations of urine culture could verify the results of NTS detection on admission in 5 cases (1.53%). NTS in 2 cases (0.61%) could cover the uropathogens of subsequent several urine cultures. Conclusions:NTS has the advantages of rapid, sensitive and comprehensive detection of urinary tract infection pathogens. When urine culture is not yet reported or even negative, NTS already has a certain clinical reference value and can be used as an effective supplement to urine culture, which is conducive to the comprehensive judgment of the patient's condition.

Objective:To compare the performance and surgical outcomes of domestic single-use digital flexible ureteroscopes with reusable digital flexible ureteroscopes in treatment of upper urinary stones.Methods:A prospective, single-blind, multicenter and randomized controlled study was performed from September 2018 to June 2019. Eligible patients were randomly assigned, in a ratio of 1∶1, to either experimental group or control group. The inclusion criteria for the study were: aged 18-75 years, solitary upper urinary stone with stone size between 0.8 and 2.0 cm and CT value less than 1 400 HU, negative preoperative urine culture and normal renal function. Exclusion criteria included: patients with acute urinary tract infection, intransitable urethral strictures, impassable ureteropelvic junction obstructions, systemic hemorrhagic disease, coagulation function abnormalities or bleeding tendency, severe hypertension or cardiopulmonary insufficiency, severe hip malformation and difficulty in meeting the demand of operation position and pregnant and lactation women. The device used in the experimental group was a domestic single-use digital flexible ureteroscope, and the device used in the control group was an imported Olympus digital flexible ureteroscope. The qualified rate of clinical comprehensive evaluation (including image quality and operational performance), the rate of device failure, the stone-free rate and the occurrence rate of adverse events (including increase in urine red blood cell and white blood cell counts, postoperative hematuria, nausea, vomiting, dizziness, and fever) in the two groups were recorded.Results:A total of 186 eligible study cases were collected from the People's Hospital of Wuhan University, the First Affiliated Hospital of Xiamen University, and the First Affiliated Hospital of Guangzhou Medical University. 90 cases in the final experimental group and 88 cases in the control group completed the trial and were included in the evaluation. There were no statistically significant differences among age [(48.40±11.36) vs. (47.40±12.53)years old, P=0.594], male to female ratio (62/28 vs. 56/32, P =0.874), BMI [(24.8±2.1) kg/m 2 vs. (25.1±2.0)kg/m 2,P =0.331], hydronephrosis (no/slight vs. mild/severe) (62/28 vs. 65/23, P =0.874), stone location and stone size [(12.8±4.7) mm vs. (11.9±5.2) mm, P =0.227]. There were no significant differences in terms of qualified rate of clinical comprehensive evaluation [98.9% (89/90) vs. 100.0% (88/88), P =0.991], lithotripsy success rate [84.4% (76/90) vs. 84.1% (74/88), P =0.888], device failure/defect rate (both 0%), and the incidence of adverse events [50.0% (45/90) vs. 52.0% (51/88), P =0.894]. The highest incidence of adverse events in two groups was the increase of red blood cells and white blood cells of routine urine after operation. There was no serious adverse event in the experimental group and 1 serious adverse event in the control group. Conclusions:There was no significant difference in image quality, device failure/defect rate, lithotripsy success rate, and adverse event rate between single-use digital flexible ureteroscopes and reusable digital flexible ureteroscopes for lithotripsy of upper ureteral and pelvic stones. Domestic single-use digital flexible ureteroscopes have good safety and effectiveness in the treatment and microscopy of upper urinary tract stones.

Objective:To explore the efficacy and safety of robot assisted laparoscopic sacrocolpopexy in the treatment of pelvic organ prolapse.Methods:The data of 16 patients who underwent robot assisted laparoscopic sacrocolpopexy in Beijing Hospital from September 2019 to May 2022 were collected. The average age was (73.5±9.3) years, the preoperative course was 4-240 months, the body mass index was (24.2±1.7) kg/m 2, the number of births was (1.7±0.8), and the quantitative index of pelvic organ prolapse (POP-Q) was grade Ⅲ -Ⅳ. The maximum urine flow rate before operation was (9.6±3.4) ml/s, the maximum detrusor pressure during urination was 26 (20, 32) cmH 2O, the first sense urine volume of the bladder was (119.1±39.4) ml, the first sense urine urgency volume of the bladder was (253.6±75.7) ml, the maximum bladder pressure capacity was (406.0±79.8) ml, and the residual urine volume was 10 (10, 28) ml. The preoperative PFDI-20 score was 100 (70, 122) and the PFIQ-7 score was 107 (90, 160). During the robot assisted laparoscopic sacrocolpopexy, the right area of the sacral promontory was exposed, the anterior and posterior walls of the uterus were separated, and the 2 cm small hole was separated from the right broad ligament of the uterus. The mesh was cut into a "Y" shape and passed through the small hole. The anterior and posterior leaves of the Y-shaped mesh were sutured to fix the cervix, and the other end was fixed to the anterior longitudinal ligament of the sacrum. The operation time and intraoperative bleeding were observed. The effect of surgery was evaluated by preoperative and postoperative urodynamic imaging, POP-Q stage, PFDI-20 score and PFIQ -7 score. Results:All the 16 operations were successfully completed. No injury of urethra, bladder, rectum, important blood vessels and nerves occurred during the operation. The average operation time was (255.6±56.0) min, and the average amount of bleeding was (28.8±18.2) ml. There was no inhibitory contraction wave in 7 patients (44%) before operation, suggesting that there was detrusor overactivity. After operation, the detrusor overactivity disappeared or significantly decreased in 7 patients. The postoperative follow-up period was 3-36 months. During the follow-up period, one patient had recurrence, and the rest had no prolapse and urination problems. The POP-Q stage was reduced to grade 0-Ⅰ after the operation. The subjective satisfaction rate of patients was 94%. The PFDI-20 score [13(8, 24)] and PFIQ -7 score [11(6, 15)] after operation were significantly reduced ( P<0.001), the initial urgent volume of bladder (272.5±88.5) ml was significantly increased compared with that before operation ( P=0.038), and the maximum volume of bladder (427.2±79.2) ml was significantly increased compared with that before operation ( P=0.006). Image urodynamics showed that the patient basically recovered the pelvic floor anatomy and achieved functional reduction. Conclusions:Robot assisted laparoscopic sacrocolpopexy has good subjective and objective effects in POP, low recurrence rate and less complications. It needs a larger sample size study for confirming the improvement of bladder function.

Objective:To investigate the availability and safety of a domestic disposable digital flexible cystoscope compared with a reusable Olympus digital flexible cystoscope in cystoscopy and removal of double J stent.Methods:From August 2018 to March 2019, patients were enrolled in this prospective, open, multicenter, randomized, parallel positive controlled clinical trial study, which were from department of Urology in Renmin Hospital of Wuhan University, the First Affiliated Hospital of Xiamen University and the First Affiliated Hospital of Guangzhou Medical University. The experimental group and control group were assigned into a 1∶1 ratio by random table method. Inclusion criteria included age≥18 years and have indications for cystoscopy or removal of double J stent. Exclusion criteria included patients having acute genitourinary tract infection, having tuberculous bladder contracture, bladder capacity less than 50ml, having urethrostenosis, female menstrual period, pregnancy and lactation, having difficulty for lithotomy position, having serious cardio-cerebrovascular disease and liver or kidney dysfunction. A domestic disposable digital flexible cystoscope was adopted in the experimental group, whereas a reusable Olympus digital flexible cystoscope was used in the control group. Acceptability of image was defined as primary availability indicator, while success rate of working and performance score were defined as secondary availability indicators and mean operating time was calculated for cystoscopy only and cystoscopy plus removal of double J stent respectively, yet rate of adverse event as well as rate of equipment defects were sorted as safety indicators.Results:A total of 188 cases which were listed in per protocol set completed the clinical trial study successfully. There were 95 cases in the experimental group and 93 cases in the control group. Acceptability of image was 93.68%(89/95) and 96.77%(90/93) respectively in two groups( P=0.52). Success rate of working was 100.00%(95/95) and 98.92%(92/93) respectively in two groups ( P=0.49). Performance score was 14.41±0.93 and 14.56±0.84 respectively in two groups ( P=0.23). Mean operating time (MOT) only for cystoscopy was (15.3±2.6) min and (15.4±3.3)min respectively in two groups ( P=0.93), while MOT for cystoscopy plus removal of double J stent was (21.0±3.2) min and (21.7±3.9) min respectively in two groups ( P=0.69). Rate of adverse event was 8.42%(8/95) and 9.68%(9/93) respectively in two groups( P=0.76). There was no equipment defects in both groups. Conclusions:There is no statistical difference in acceptability of image, success rate of working, performance score, mean operating time for cystoscopy or removal of double J stent, rate of adverse events and rate of equipment defects. A domestic disposable digital flexible cystoscope has shown non-inferiority in the availability and safety compared with a reusable Olympus digital flexible cystoscope.

Objective:To explore the potential relationship of urethral pressure profilometry and the complications of artificial urethral sphincter (AUS) implantation.Methods:The clinical data of patients who underwent AUS implantation in Beijing Hospital from March 2019 to March 2022 were retrospectively analyzed. All the patients were male. The average course of disease was 43.1 months(ranging 11-120 months). The average age was (68.6±13.2) years. The median number of pads used was 4.5 (3.0, 6.0). The preoperative maximum urethral pressure (MUP) was (84.6±25.5) cmH 2O, and the maximum urethral closure pressure (MUCP) was 51.0 (41.0, 74.0) cmH 2O. AUS implantation was performed through a single perineal incision in all patients. The sleeve size was mainly determined by the measured urethral circumference of the patient. After installation of all components, the urethral pressure profilometry was performed under the state of device inactivation and activation. The pump was activated 6 weeks after the operation, and telephone follow-up was performed 3 months after the activation of the device. The urinary control and complications were recorded. The results of follow-up were compared with the results of urethral pressure profilometry, and the preliminary conclusions were drawn. Results:In this study, 3 patients (20%) received 4.0cm cuffs, 10 patients (66.7%) received 4.5cm cuffs, and 2 patients (13.3%) received 5.0cm cuffs. The MUP and MUCP of AUS device in inactivated state were (82.5±30.2) cmH 2O and 51.0(48.0, 77.0) cmH 2O. In the activated state, MUP was (138.9±21.7) cmH 2O and MUCP was 109.0(94.0, 133.0) cmH 2O. Compared with that before operation, the urethral pressure in the inactivated state did not increase significantly (all P > 0.05), while the urethral pressure in the activated state increased significantly (all P < 0.001). The patients were followed-up for 3-33 months. Thirteen patients (86.7%) used the initial installation device, and all of them met the standard of social continence. One patient died of cerebrovascular accident. One patient took out the device due to urethral erosion. The incidence of complications was 26.7% (4/15), including painless hematuria in 2 cases, scrotum and penis infection in 1 case, and urethral erosion in 1 case. The MUP and MUCP of these patients were (100.0 ± 40.7) cmH 2O and (80.8 ± 39.7) cmH 2O respectively. In the intraoperative active state, the MUP was (151.5 ± 15.3) cmH 2O and the MUCP was (123.0 ± 17.2) cmH 2O. The MUP of the other 3 patients in the device activation state was significantly higher than the average value, and all of them were above 150 cm H 2O, except one patient who was infected due to cognitive problems and chronic urinary retention. In 11 patients without complications, the MUP and MUCP were (76.1±24.7) cmH 2O and (55.1±20.0) cmH 2O respectively. In the intraoperative active state, the MUP was (134.4±22.5) cmH 2O and the MUCP was (108.5±29.8) cmH 2O. Conclusions:AUS implantation has a definite curative effect. Poor comprehension, and MUP higher than 150 cmH 2O in the activated state of the device may be risk factors for complications.

Triterpenoids are a class of natural products of great commercial value that are widely used in pharmaceutical, health care and cosmetic industries. The biosynthesis of triterpenoids relies on the efficient synthesis of squalene epoxide, which is synthesized from the NADPH dependent oxidation of squalene catalyzed by squalene epoxidase. We screened squalene epoxidases derived from different species, and found the truncated squalene epoxidase from Rattus norvegicus (RnSETC) showed the highest activity in engineered Escherichia coli. Further examination of the effect of endogenous cytochrome P450 reductase like (CPRL) proteins showed that overexpression of NADH: quinone oxidoreductase (WrbA) under Lac promoter in a medium-copy number plasmid increased the production of squalene epoxide by nearly 2.5 folds. These results demonstrated that the constructed pathway led to the production of squalene epoxide, an important precursor for the biosynthesis of triterpenoids.
Animals ; Escherichia coli/genetics* ; NADPH-Ferrihemoprotein Reductase ; Protein Engineering ; Rats ; Repressor Proteins ; Squalene ; Squalene Monooxygenase/genetics*

Objective:To evaluate the efficacy and safety of robotic arm assisted laparoscopic hysterosacral fixation in patients with pelvic organ prolapse(POP), and its impact on lower urinary tract function.Methods:This study retrospectively analyzed the clinical data of POP patients who had undergone robotic arm assisted laparoscopic hysterosacral fixation at our center from June 2019 to October 2020, and conducted exploratory research.Results:A total of 6 patients were included in the study, with POP quantitative staging above stage Ⅲ.The ages ranged from 70 to 82 years.The number of births each patient had given ranged from 1 to 3, & all were via vaginal deliveries.There were no significant changes in urodynamic parameter scores in any patients before and after surgery, but half of the patients had detrusor overactivity before surgery, which all disappeared after surgery.In addition, synchronous X-ray images showed that the postoperative pelvic organs were closer to the normal anatomical position.At the same time, quantitative staging of POP had achieved clear improvement, and related scale scores also significantly improved.One patient complained of occasional lumbar and back discomfort with postoperative over-stretching during outpatient review, which improved after symptomatic treatment.Conclusions:robotic arm assisted Laparoscopic hysterosacral fixation is satisfactory in efficacy and safety for POP patients, with good postoperative restoration of the uterus to the anatomical position and has insignificant influence on the function of the lower urinary tract.It is worth further assessment for wide application.

Objective A retrospective study was conducted to investigate the clinical epidemiology of ufinary calculus in Hubei province to provide the population characteristics,stone distribution characteristics,treatment methods,surgical costs and other clinical epidemiological data of patients with trinary calculus,and to provide clinical evidence for the development of prevention and treatment measures for urinary calculi.Methods According to the geographical position,local tertiary hospitals were selected as survey objects and medical records such as age,sex,occupation,stone position,type of operation,and cost of surgical treatment of urinary stone disease were retrospectively investigated between January,2015 and December,2017.Result A total of 193 784 patients were enrolled in this study from 27 hospitals in Hubei province.In which,76 209 were admitted to the hospital for urinary calculus,accounting for 39.33％.Among them,49 460 patients were males (64.90％) and 26 749 were females(35.10％),which the ratio was 1.85∶ 1.All age groups can suffer from urinary calculus,with the peak age range from 41 to 70 years old with 58 985 cases,accounting for 77.40％.The main proportion of patients with urinary calculus were farmers with 21 818 cases (28.63％),comparing with workers (9.74％),busioess man (5.58％),government employees (5.14％),retirees (11.25％) and unemployed (12.58％).A total of 71 439 patients with upper urinary calculus and 4 770 patients with lower urinary calculus and the ratio of upper and lower urinary calculus was 14.98∶ 1.From 2015 to 2017,23 567 patients with urinary calculi underwent ureteroscopic]ithotripsy,and 14 681 patients underwent percutaneous nephrolithotomy.The average operation cost of patients with urolithiasis was 18 395.52 RMB,18 566.31 RMB,19 080.91 RMB in 2015,2016,2017 respectively.Conclusion Hubei province is an area with high incidence of urinary calculus,accounting for 39.33％.The high incidence age is 41-70 years.The occupation composition is mainly farmers.In the past three years,the cost of surgery gradually increased.