Objective To investigate the association between expression of the epithelial?to?mesenchymal transition(EMT)biomarkers and the malignant progression of gastric cancer in primary tumors and metastases and their possible correlation with progression of gastric cancer(GC). Methods The EMT biomarkers including E?cadherin,β?catenin,N?cadherin,Snail and TGF?β1 were detected by immunohistochemical method for 145 cases of gastric cancer(GC),25 cases of abnormal hyperplasia,13 cases of intestinal metaplasia,42 cases of lymph node metastasis and 40 cases of normal gastric mucosa tissues. Results Positive rates of TGF?β1,Snail,E?cadherin,β?catenin and N?cadherin were 73.5%,65.5%, 14.5%,53.1%and 35.9%,respectively,in gastric cancer tissues and 100%,100%,0%,27.5%and 2.5%,respectively,in normal gastric tissues, with a significant difference between the two groups(P<0.05). The decreased expression of E?cadherin andβ?catenin and the increased expression of TGF?β1 were related to the depth of invasion of gastric cancer(P<0.05). The expression of E?cadherin was correlated positively with the expres?sion ofβ?catenin,but negatively with the expression of TGF?β1. Whereas,the expression of N?cadherin was correlated positively with the expression of TGF?β1(P<0.05). The expression of E?cadherin andβ?catenin in lymph node metastasis was significantly higher than that in gastric cancer tis?sues,while the expression of TGF?β1 was lower than in gastric cancer tissues(P<0.05). Conclusion The increased expression of TGF?β1 and Snail and the decreased expression of E?cadherin,β?catenin,and N?cadherin are involved in the processes of invasion and metastasis of GC. The transformation of E?cadherin to N?cadherin and the expression of TGF?β1 may play an important role in the development of GC. In lymph node me?tastasis,the phenomenon of mesenchymal?to?epithelial transition(MET)occurs.

BACKGROUND:Urothelial cells are important seeding cells for urinary tissue engineering, but they are difficult to proliferate in vitro. Several studies have shown that bone marrow mesenchymal stem cells can differentiate into urothelial cells, but how these cells functions in vivo in epithelium generation after implantation, and the application of these cells in tissue engineering, are rarely studied. OBJECTIVE:To explore the isolation and proliferation of rabbit bone marrow mesenchymal stem cells that are induced into urothelial cells in combination with rabbit bladder acellular matrix to construct tissue-engineered grafts, and to assess the effect of the induced cells as seeding cells. METHODS:Twelve 8-week-old male New Zealand white rabbits were chosen to obtain bone marrow samples through tibia puncture, and to isolate bone marrow mesenchymal stem cells by density gradient centrifugation. Then the fourth or fifth generation of bone marrow mesenchymal stem cells were cultured in conditioned medium for 2 weeks, and then identified by PCR and immunofluorescence. After that, the induced cells were seeded on rabbit bladder acellular matrix to construct tissue-engineered grafts for bladder repairing. Another 12 rabbits served as control group, and urothelial cells combined with bladder acellular matrix was used for bladder repairing. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells were successful y cultured and proliferated in vitro. After induction, PCR detection suggested that stem cellmarker (CD44) expression decreased, and epithelial cellmarker (UP1a) expression increased in the induced cells. Immunofluorescence staining demonstrated that the induced cells rather than bone marrow mesenchymal stem cells were positive for specific urothelial marker, UP1a. A stable continuous epithelial layer was observed on tissue-engineered grafts constructed by induced cells after 2 weeks, similar to the grafts built by urothelial cells. Induced bone marrow mesenchymal stem cells can differentiate into urothelial cells that can be used as seeding cells for urinary tissue engineering, which may be another choice out of urothelial cells.

BACKGROUND:The main way for long-segmental ureteral reconstruction may cause a lot of traumas and complications. Therefore, to seek a new repair method is urgent. OBJECTIVE:To investigate the feasibility of a tissue-engineered tubular graft for ureteral reconstruction. METHODS:Bone marrow mesenchymal stem cels and smooth muscle cels of rabbits were seeded into the two surfaces of bladder acelular matrix and cultivated for 7 days. Then the graft was used to prepare a 4-cm long tissue-engineered tubular graft, which was regarded as experimental group. Smooth muscle cels seeded onto the bladder acelular matrix was used to construct the tissue-engineered tubular graft as control group. Twenty-five New Zealand rabbits were randomly divided into experimental group (n=20) and control group (n=5), and two kinds of tubular grafts covered with omentum were implanted into the two groups, respectively, for repair of ureteral defects. Hematoxylin-eosin staining and immunohistochemical detection were performed at 2, 4, 8 weeks after implantation. RESULTS AND CONCLUSION:In the experimental group, hematoxylin-eosin staining showed epithelial coverage and muscle fibers on the lumen of tissue-engineered tubular grafts at 8 weeks after implantation; immunohistochemistry showed that anti-AE1/AE3 antibody and anti-uroplakinⅢa antibody were positive, confirming that there were mature epithelial cels on the lumen of tissue-engineered tubular grafts. In the control group, five rabbits were dead within 2 weeks after removal of ureteral scaffold, and autopsy showed scar formation inside the graft and severe hydronephrosis. These results demonstrate that it is feasible to construct the tissue-engineered tubular graft using bone marrow mesenchymal stem cels and smooth muscle cels into the bladder acelular matrix for ureteral reconstruction. Bone marrow mesenchymal stem cels can potentialy promote urothelial regeneration.

Objective To evaluate the safety and efficacy of ultrasound guided flexible ureteroscopic lithotripsy.Methods From January 2015 to December 2016,a randomized clinical trial enlisted 130 patients,presenting symptomatic renal stones with 1.3-2.0 cm,in our center.Patients were randomly assigned to ultrasound or radical guided retrograde intrarenal surgery (RIRS) with flexible ureteroscope.There were 37 male and 28 female patients in ultrasound group.Their mean age,BMI index and stone size were (47.7 ± 21.3) years,(22.1 ± 4.7) kg/m2 and (1.4 ± 0.2) cm,respectively.In radical group,there were 22 male and 43 female patients.Their mean age,BMI index and stone size were (51.3 ± 19.2) years,(21.6 ± 3.3) kg/m2 and (1.6 ± 0.1) cm,respectively.There was no significant difference of those items between ultrasound and radical group.In ultrasound group,the transducer was placed at the dorsal and ventral to visualize the real time image of ipsilateral renal collecting system.The safety wire placing,ureteroscope passing through the uretheral access sheath and the lithotripsy detail could be easily monitored.We compared stone size,operative time,stone-free status and complication rates between the ultrasound and radical group.Results All patients accepted the operation successfully.Operative time was significantly shorter in the ultrasound group than that in radical group.[(62.4 ± 31.7) min vs.(80.4 ± 42.1)min,P ＜0.05].The average radical exposure during was (37.5 ± 25.2)seconds in radical group.Ureteral injury was seen in 20.0％ (13/65),18.5％ (12/65) cases in ultrasound and radical group,respectively.The mucosal injury rate in each group was 92.3％ (12/13) and 83.3％ (10/12) (P ＞ 0.05),respectively.Muscle layer injury was recorded in 1 case of in ultrasound group and 2 cases in radical group.Clavien grade Ⅳ or higher grade complications was not observed in both groups.Fever,pain,nausea or vomiting,persistent hematuria in ultrasound group were 2 cases,3 cases,1 cases and 5 cases.In radical group,those complications were in 2 cases,3 cases,7 cases and 1 cases,respectively.There was no significant difference between the two groups (P ＞ 0.05).The ultrasound group showed no significant difference in the stone-free rate [90.8％ (118/130)vs.89.2％ (116/130)] or the complication rate compared to those in radical group(P ＞ 0.05).Conclusions The retrograde intra-renal lithotripsy guided solely by ultrasound can be carried out in the general population in an effective and safe fashion without ionizing radiation,which has the potential to replace radical guided in flexible ureteroseopy.

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.

INTRODUCTIONAccording to the findings of some studies, instability due to inertia during changes in speed may negatively impact the quality of chest compressions performed during cardiopulmonary resuscitation (CPR) in a moving environment. This study thus aimed to introduce a simple device that maintains the balance of a person performing CPR in a moving environment, such as an ambulance. We also sought to evaluate the effectiveness of this device in the improvement of the quality of chest compressions.

METHODSThe experiment comprised a total of 40 simulated cardiopulmonary arrest scenes (20 in the experimental group and 20 in the control), in which CPR was conducted by eight paramedics. Each simulation involved two paramedics randomly selected from the eight. The ambulance took the same route from the simulated site to the hospital, and continuous CPR was performed on a manikin in the ambulance with or without the aid of our proposed novel device.

RESULTSThe average number of chest compressions per simulation in the experimental and control groups was 1330.75 and 1266.60, respectively (p = 0.095). The percentage of chest compressions with adequate depth achieved in the experimental and control groups was 72% ± 4% and 50% ± 3%, respectively (p < 0.0001).

CONCLUSIONBy maintaining the balance of the CPR performer, our proposed novel device can offset the negative impact that instability (due to a moving environment) has on chest compressions. The device may also lead to an increase in the percentage of chest compressions that achieve adequate depth.

Adult ; Allied Health Personnel ; Ambulances ; Cardiopulmonary Resuscitation ; instrumentation ; methods ; Emergency Medical Services ; methods ; Equipment Design ; Equipment Safety ; Female ; Humans ; Male ; Manikins ; Movement ; Out-of-Hospital Cardiac Arrest ; mortality ; therapy ; Physical Exertion ; physiology ; Reference Values ; Risk Assessment ; Sensitivity and Specificity ; Survival Rate ; Treatment Outcome

Objective:To identify the risk factors associated with postoperative adjuvant chemotherapy in patients with stage I gastric cancer and establish nomograms model based on risk factors.Methods:In this retrospective case-control study, 161 cases with stage Ⅰ primary gastric adenocarcinoma were included who underwent gastrectomy at the Department of General Surgery of the First Medical Center of Chinese PLA General Hospital from January to December in 2020, including 129 male cases and 32 females cases, with the average age of (59.90±0.80) years. Among them, 41 cases were treated with postoperative adjuvant chemotherapy (chemotherapy group), while 120 cases who did not receive postoperative adjuvant chemotherapy (no chemotherapy group). Univariate and multivariate Logistic regression analyses were used to identify the risk factors of adjuvant chemotherapy in stage Ⅰ gastric cancer patients and establish the nomograms predictive model. ROC curve and calibration curve were used to evaluate the performance of the model.Results:Multivariate analysis revealed that primary tumor site, tumor size, T stage, N stage lymph-vascular tumor embolus or perineural invasion were the independent risk factors of postoperative adjuvant chemotherapy for stage Ⅰ gastric cancer( P<0.05). The ROC curve indicated that area under the curve (AUC) of the multivariate model was 0.91(95% CI: 0.86-0.97). The calibration curve showed that probability predicted by nomograms was consistent with the actual situation(C-index: 0.91). Conclusions:The tumor located in the proximal stomach, tumor size>2 cm, T 2, N 1, lymph-vascular tumor embolus or perineural invasion maybe be the risk factors for chemotherapy decision in stage Ⅰ gastric cancer patients. The established model has good predictive ability for postoperative chemotherapy of stage Ⅰ gastric cancer patients, which might provide reference for the selection of clinical decisions in this part of patients.

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 investigate clinicopathological features of carcinoma showing thymus-like elements (CASTLE) in the head and neck regions. Methods: Clinicopathological data of 7 patients with CASTLE in the head and neck regions were retrospectively reviewed.Immunohistochemical staining and in situ hybridization for EBER were performed. BRAF^V600E mutation was examined by ARMS method in 6 cases. Results: There were 5 females and 2 males with age between 49 and 78 years (average of 65.6 years). All tumors were solitary nodular lesions with an infiltrative border, including 6 intrathyroid tumors and 1 extrathyroid tumor in the laryngeal pharynx.The tumors were 1.7-4.1 cm in diameter (average of 3.0 cm). Four cases demonstrated lymph node metastasis.All patients were alive without metastasis during follow-up, except one consultation case (with FNA sample) developed recurrence at the primary site. The cases showed different immunoreaction to CD5, diffuse immunoreaction with p63, CK5/6 and CD117, but negative staining for TTF1, TG and calcitonin. One case showed positive immunoreaction with Synin less than 30% tumor cells. The Ki-67 labeling index was between 3% and 90%. No BRAF^V600E mutation and EB virus infection were detected. Conclusions Extrathyroid CASTLE involving laryngeal pharynx shows the similar morphological and immunohistochemical features with intrathyroid CASTLE.Immunohistochemical markers of CD5 and CD117 are helpful in the diagnosis. Ki-67 labeling index can be high in CASTLE, especially in lymphoepithelioma type. CD5-negative CASTLE may have neuroendocrine differentiation. BRAF^V600E mutation and EB virus may not be involved in the carcinogenesis of CASTLE.

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.